The wheat NLR pair RXL/Pm5e confers resistance to powdery mildew

Powdery mildew poses a significant threat to global wheat production and most cloned and deployed resistance genes for wheat breeding encode nucleotide-binding and leucine-rich repeat (NLR) immune receptors. Although two genetically linked NLRs function together as an NLR pair have been reported in other species, this phenomenon has been relatively less studied in wheat. Here, we demonstrate that two tightly linked NLR genes, RXL and Pm5e, arranged in a head-to-head orientation, function together as an NLR pair to mediate powdery mildew resistance in wheat. The resistance function of the RXL/Pm5e pair is validated by mutagenesis, gene silencing, and gene-editing assays. Interestingly, both RXL and Pm5e encode atypical NLRs, with RXL possessing a truncated NB-ARC (nucleotide binding adaptor shared by APAF-1, plant R proteins and CED-4) domain and Pm5e featuring an atypical coiled-coil (CC) domain. Notably, RXL and Pm5e lack an integrated domain associated with effector recognition found in all previously reported NLR pairs. Additionally, RXL and Pm5e exhibit a preference for forming hetero-complexes rather than homo-complexes, highlighting their cooperative role in disease resistance. We further show that the CC domain of Pm5e specifically suppresses the hypersensitive response induced by the CC domain of RXL through competitive interaction, revealing regulatory mechanisms within this NLR pair. Our study sheds light on the molecular mechanism underlying RXL/Pm5e-mediated powdery mildew resistance and provides a new example of an NLR pair in wheat disease resistance.